Thermo Scientific™ Eco47III (AfeI)
The Eco47III (AfeI) restriction enzyme recognizes AGC^GCT sites and cuts best at 37°C in O buffer (Isoschizomers: AfeI, Aor51HI).
Brand: Thermo Scientific™ ER0321
Code : 88
Additional Details : Weight : 0.01000kg
5'...A G C▵G C T...3'
3'...T C G▵C G A...5'
Conditions for 100% Activity:
- 1X Buffer O:50mM Tris-HCl (pH 7.5 at 37°C), 10mM MgCl2, 100mM NaCl and 0.1mg/mL BSA
- Incubate at 37°C
- Eco47III is supplied in:10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with Eco47III, more than 80% of pBR322 DNA fragments can be ligated and more than 90% of these can be recut
- Dam: never overlaps-no effect
- Dcm: never overlaps-no effect
- CpG: completely overlaps-blocked
- EcoKI: never overlaps-no effect
- EcoBI: may overlap-effect not determined
Digestion of Agarose-embedded DNA:
- Minimum 5U of the enzyme are required for complete digestion of 1μg of agarose-embedded lambda DNA in 16 hours